Cov khoom

Ib leeg- nucleus RNA Sequencing

Kev nce qib hauv tib lub xov tooj ntawm tes tuav thiab ib tus neeg siv tsev qiv ntawv tsim cov txheej txheem sib txuas nrog kev sib txuas ua haujlwm siab tso cai rau cov kev tshawb fawb gene ntawm cov xov tooj ntawm tes.Nws ua kom muaj kev sib sib zog nqus thiab ua tiav kev tshuaj xyuas ntawm cov cell complex, nyob rau hauv uas nws feem ntau zam qhov npog ntawm lawv cov heterogeneity los ntawm kev noj nruab nrab ntawm tag nrho cov hlwb.

Txawm li cas los xij, qee lub hlwb tsis tsim nyog los ua rau hauv ib leeg-cell ncua kev kawm, yog li lwm cov qauv kev npaj - xav tau kev rho tawm nucleus los ntawm cov ntaub so ntswg, uas yog, nucleus raug rho tawm ncaj qha los ntawm cov ntaub so ntswg lossis cell thiab npaj rau hauv ib qho kev ncua rau ib leeg- cell sequencing.

BMK muab 10 × Genomics ChromiumTM raws li ib leeg-cell RNA sequencing kev pabcuam.Cov kev pabcuam no tau siv dav hauv kev tshawb fawb txog kev tshawb fawb txog kab mob, xws li kev tiv thaiv kab mob sib txawv, qog heterogeneity, kev loj hlob ntawm cov ntaub so ntswg, thiab lwm yam.

Spatial transcriptome nti: 10 × Genomics

Platform: Illumina NovaSeq Platform

Tiv tauj peb

Cov ntsiab lus kev pabcuam

Bioinformatics

Cov txiaj ntsig Demo

Technical txoj cai

Kev sib cais ntawm nuclei yog ua tiav los ntawm 10 × Genomics ChromiumTM, uas muaj yim-channel microfluidics system nrog ob txoj kev hla.Hauv qhov system no, gel hlaws nrog barcodes thiab primer, enzymes thiab ib lub nucleus yog encapsulated nyob rau hauv nanoliter-loj roj poob, generating Gel Bead-in-Emulsion (GEM).Thaum GEM raug tsim, cell lysis thiab tso tawm barcodes tau ua nyob rau hauv txhua GEM.mRNA yog thim rov qab rau hauv cDNA molecules nrog 10 × barcodes thiab UMI, uas yog ntxiv rau cov qauv sib txuas hauv tsev qiv ntawv.

Cov ntaub so ntswg tsis haum rau kev npaj ib leeg ntawm tes

Cell / daim ntaub	Yog vim li cas
Unfresh khov cov ntaub so ntswg	Tsis tuaj yeem tau txais cov koom haum tshiab lossis kev txuag ntev
Muscle cell, Megakaryocyte, Fat…	Cell txoj kab uas hla yog loj dhau mus rau hauv lub ntsuas
Lub siab…	Yooj yim heev rau tawg, tsis tuaj yeem paub qhov txawv ntawm ib leeg
Neuron cell, Lub hlwb…	Ntau rhiab heev, yooj yim rau kev ntxhov siab, yuav hloov cov txiaj ntsig sib luag
Pancreas, thyroid…	nplua nuj nyob rau hauv endogenous enzymes, cuam tshuam rau zus tau tej cov cell suspension

Single-nucleus vs Single-cell

Ib leeg-nucleus	Ib leeg xwb
Unlimited cell diam	Cell txoj kab uas hla: 10-40 μm
Cov khoom tuaj yeem yog cov ntaub so ntswg khov	Cov khoom yuav tsum yog cov ntaub so ntswg tshiab
Tsawg kev ntxhov siab ntawm cov cell khov	Kev kho enzyme tuaj yeem ua rau muaj kev ntxhov siab ntawm tes
Tsis muaj cov qe ntshav liab yuav tsum tau muab tshem tawm	Cov qe ntshav liab yuav tsum tau muab tshem tawm
Nuclear qhia txog bioinformation	Tag nrho cov cell qhia txog bioinformation